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Author Topic: Dihydroethidium Staining on Brain Sections  (Read 5065 times)

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Kaushik.shah

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Dihydroethidium Staining on Brain Sections
« on: September 15, 2008, 10:45:50 PM »
Hi All!

I have to use the unfixed mouse brain sections for Dihydroethidium (DHE) microscopy to see the Reactive Oxygen Species, superoxide generation.

I have a problem with frozen unfixed Brain sections (snap freeze in isopentane/dryice slurry), which get loose when I incubate them at 37 degree in humidified chamber after collecting sections on to the slides. it really doesnt stick to the slides. I use a Fisher Superfrost slides. and I keep the slides  all the time in dryice until process it for DHE staining.

Can anyone suggest me the Mouse brain processing protocol that can be used for Dihydroethidium staining. I mean detailing on sectioning and using the kind of slides or any other way of having unfixed brain sections for DHE fluorescence Microscopy.

thanks

Dihydroethidium Staining on Brain Sections
« on: September 15, 2008, 10:45:50 PM »

Offline richard03

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Re: Dihydroethidium Staining on Brain Sections
« Reply #1 on: November 23, 2008, 11:19:30 PM »
The unfixed tissue sections must be air dried for at least 30 minutes before any staining procedure. Use charged slides or gelatin coated slides should help as well.

Re: Dihydroethidium Staining on Brain Sections
« Reply #1 on: November 23, 2008, 11:19:30 PM »